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. 2015 Nov 9;48(1):45–54. doi: 10.3892/ijo.2015.3237

Figure 7.

Figure 7

Morphological changes in PANC-1 cells after treatment with GEF and macrolides. (A) May-Giemsa staining: May-Giemsa staining was performed after treatment with either GEF (25 μM) or gemcitabine (100 μM) in the presence or absence of EM900 (50 μM) for 48 h. Scale bars, 10 μm. Arrows indicate nuclear chromatin condensations. (B) Electron microscopy: PANC-1 cells were processed for electron microscopy after treatment with GEF (25 μM), AZM (50 μM), GEF+AZM, and gemcitabine (100 μM) for 48 h. Lower panels are at higher magnifications of the squared areas in each upper panel. Scale bars, 2 μm. (C) PANC-1 cells were treated with GEF (25 μM), AZM (50 μM), EM900 (50 μM), GEF+AZM, and gemcitabine (10 and 100 μM) for 48 h. Cellular proteins were separated by 15% SDS-PAGE for caspase-3 and 7.5% for PARP and immunoblotted with either anti-cleaved caspase-3 Ab or anti-PARP Ab. Cell lysate derived from HL-60 cells treated with vitamin K2 was used as a positive control for apoptosis induction (42). (D) PANC-1 cells were treated with various concentrations of either necrostatin-1 or Z-VAD-FMK in the presence or absence of GEF (50 μM), GEF (50 μM) plus EM900 (50 μM), or gem (300 μM) for 48 h. The number of viable cells was assessed using CellTiter Blue.